PROJECT SUMMARY We are moving into the era of precision medicine of cancer, where the sequencing of tumors in patients will better inform clinical decisions. However, the advancement of precision medicine will require the distinction of those mutations that causally drive the disease from an ocean of bystander mutations. It will be equally important to identify novel regulatory functional sites in proteins that serve as targets for novel therapeutic drugs. The main goal of my proposal is to identify functional sites in protein kinases (a subset of proteins highly mutated in cancer), so that we enable the identification of cancer driver mutations and novel ?druggable? sites. To achieve this goal, I propose to deploy a wide range of technologies, including computational models, mass-spectrometry, biochemical, cellular and in vivo screens of protein function, that I am currently developing in collaboration with experts in these fields. In aim 1, I propose to develop and validate a suite of novel computational models that will allow the identification of functional sites that are either common or unique to distinct kinase groups. My preliminary data suggests that these comparative coupling models can not only accurately recapitulate known functional distinctions between kinases, but also pinpoint uncharacterized functional sites that appear to be unique to specific kinase groups. In aim 2, I propose to experimentally uncover several novel regulatory sites using the allosterically-regulated kinase Fus3 as a model system. As an example, our preliminary data describes the identification of two such novel regulatory sites. In aim 3, I propose to investigate how cancer mutations perturb kinase function and cellular survival by studying a specific mutation that is observed in leukemia patients that develop resistance to Abl inhibitors. Our preliminary data indicates that this mutation leads to changes in kinase substrate specificity that may then deregulate cell cycle progression. In addition to my formal advisor (Prof. Michael Yaffe), these projects have allowed me to establish a network of high-caliber career mentors (Profs. Rama Ranganathan, Forest White and Michael Hemann) and collaborators (Prof. Chris Bakal and David Pincus). These mentors have given me increasing independence, as proven by having launched projects in areas unexplored by them, the acquisition of competitive postdoctoral funding or the establishment of new collaborations. Moreover, they will generously allow me to take my research with me as I transition to independence. With their guidance, I have designed a comprehensive training plan that will boost my prospects of securing a faculty position at a top research institution. In addition to training in grant- writing, teaching and leadership skills, this plan includes learning X-Ray crystallography, a technique that we have identified as critical for my future success. Thanks to this plan and the excellent research environment provided by my mentoring team, the Koch and MIT, I enthusiastically look forward to initiating a successful research program studying the mechanism of action of cancer mutations.